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Chaudhary Charan Singh Haryana Agricultural University, Hisar
Chaudhary Charan Singh Haryana Agricultural University popularly known as HAU, is one of Asia's biggest agricultural universities, located at Hisar in the Indian state of Haryana. It is named after India's seventh Prime Minister, Chaudhary Charan Singh. It is a leader in agricultural research in India and contributed significantly to Green Revolution and White Revolution in India in the 1960s and 70s. It has a very large campus and has several research centres throughout the state. It won the Indian Council of Agricultural Research's Award for the Best Institute in 1997. HAU was initially a campus of Punjab Agricultural University, Ludhiana. After the formation of Haryana in 1966, it became an autonomous institution on February 2, 1970 through a Presidential Ordinance, later ratified as Haryana and Punjab Agricultural Universities Act, 1970, passed by the Lok Sabha on March 29, 1970. A. L. Fletcher, the first Vice-Chancellor of the university, was instrumental in its initial growth.
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ThesisItem Open Access Immobilization of xylanase from Bacillus pumilus over aluminum oxide beads(CCSHAU, Hisar, 2021-09) Tanwar, Ekta; Nagar, SushilIn present investigation the xylanase from Bacillus pumilus SV-85S was covalently immobilized over aluminum oxide beads. The immobilization yield and efficiency were 48.05 and 83.13%, respectively. Some surface modifications were observed through SEM analysis on xylanase bound aluminum oxide beads. The bands at 3440 and 1635 cm-1 were observed through FTIR analysis over aluminum oxide beads, glutaraldehyde treated beads and immobilized xylanase. Optimum pH and temperature for free and immobilized enzyme were 7.0 and 55ᵒC, respectively. Increment of Km and Vmax values was observed in immobilized enzyme by around 28.96 and 34.61 % respectively. The pH, thermal and storage stability were improved after immobilization. The Immobilized xylanase retained 65 % of its initial activity even after 5 cycles of reusability. The increase in Half-life (t1/2) and D- value whereas decrease in kd value after immobilization, indicates that thermal stability of xylanase was enhanced. Thermodynamic parameters (ΔH, ΔG and ΔS) were enhanced after immobilization of xylanase. The decrease in activation energy in case of immobilized xylanase indicates that catalytic efficiency was enhanced. Hence, the present investigation concludes that immobilization of enzyme is important due to its considerable reusability and stability, and advantageous for industrial application.ThesisItem Open Access Comparative studies of free and immobilized Aspergillus phytase(CCSHAU, Hisar, 2021-09) Mallesh, Gampa; Nagar, SushilIn this investigation the phytase from Aspergillus oryzae SBS50 was immobilized over activated aluminum oxide beads. The immobilization yield and efficiency were 47.4 and 71.2 %, respectively. Morphology and functional groups of activated beads and immobilized phytase were determined through SEM and FTIR analysis. The immobilized phytase retained 51.5 % of its initial activity even after 5 cycles of reusability. The optimum pH and temperature were shifted from 5.5 to 4.0 and 50 to 60 C after immobilization. The Km value was 2.61 and 3.09 mM, whereas as Vmax value was 32.25 and 43.29 IU/ml for free and immobilized phytase, respectively. The pH stability, thermal stability, storage stability and thermodynamic parameters (H, G, and S) were enhanced after immobilization The increase in Half-life (t1/2) and D-value whereas decrease in Kd value after immobilization indicates that thermal stability of phytase was enhanced. Thus, the investigation concludes that immobilization of phytase causes in improvement of many properties, so it is highly beneficial to immobilize the phytase.ThesisItem Open Access In vitro antioxidant and antimicrobial activities of mustard extracts(CCSHAU, Hisar, 2021-09) Jyoti; Nisha KumariMustard is the third most important oilseed crop after oil palm and soybean. Although mustard is widely used as edible oil and condiment, the residue left after oil extraction is popularly used as livestock and poultry feed with excellent protein quality. However, the information regarding antioxidative and antimicrobial potential of mustard seed meal is still scanty. Therefore, the objective of the present study was to extract the polyphenols from brown and yellow mustard seed meal with different solvents i.e 80% methanol, 80% ethanol, 80% acetone and 80% isopropanol. Each extract was analysed for antioxidant potential using DPPH and ABTS scavenging activity, ferric ion reducing antioxidant power (FRAP) assay, total antioxidant activity, iron chelating activity and β-Carotene content. The 80% acetone extracted maximum polyphenols followed by 80% ethanol, 80% methanol and 80% isopropanol. Brown mustard exhibited a higher amount of polyphenols as compared to yellow mustard. Yellow mustard exhibited higher antioxidant potential in terms of DPPH and ABTS scavenging activity, ferric ion reducing antioxidant power, iron chelating activity and β-Carotene content. Correlation analysis indicated a negative association between total phenolic content and DPPH, ABTS and FRAP. Antimicrobial potential was evaluated against two gram-negative (Escherichia coli and Xanthomonas campesteris), two gram positive (Bacillus cereus and Lactobacillus plantarum) bacteria and three fungi (Fusarium oxysporum, Macrophomina phaseolina and Candida albicans). Yellow mustard exhibited higher antibacterial activity while brown mustard had higher antifungal activity. Both the mustard seed meal extracts were ineffective against probiotic bacteria Lactobacillus plantarum. Gallic acid, caffeic acid, ferulic acid and sinapic acid were identified in mustard seed meal by HPLC analysis.ThesisItem Open Access Effect of phytase supplementation on micronutrients dialyzability in wheat bran(CCSHAU, Hisar, 2021-09) Kiran; Vinod KumarBread wheat, a staple cereal, have considerably low amount of iron (Fe) and zinc (Zn) bioavailability mainly due to the presence of phytic acid (PA; myo-inositol-1,2,3,4,5,6-hexakisphosphate), a phosphorus storage form in seeds, as a potent chelator of divalent cations. Wheat bran (WB) is a residue of the rolled milled wheat grain and it is rich source of dietary fibres, minerals, inorganic phosphates, vitamins, and tannins. The major portion of PA is accumulated in aleurone layer and globoid crystals of wheat. The PA chelates of Fe and Zn is a major source of concern for monogastric animals as lack of suitable digestible enzyme make this complex insoluble and decreases bioavailability of these micronutrients. Supplementation of exogenous phytase for dephytinization and value addition of cereals based foods is considered as a promising approach. In present study, experimental five wheat varieties were taken and their bran samples were collected for determining their chemical and biochemical properties (including content of PA, tannin, total dietary fibre, protein, inorganic phosphate, Fe and Zn) affecting the micronutrient bioavailability by either way. Significant variation was noticed in dietary fibre, crude protein and micronutrient content. Exogenous phytases from two different sources (fungal phytase from Aspergillus orizae strain SBS50 and wheat phytase from Sigma-Aldrich) were used to treat the wheat bran samples and determine their effects on above parameters. Phytase untreated samples were taken for comparative analysis of results obtained. For phytase treatments, 0.2 U/g phytase enzyme was used for both enzyme and results revealed that the PA content and tannin content was reduced by phytase treatment in the range of 37.2-39.5 % with wheat phytase and 7.90-33.33 % with Aspergillus phytase, respectively. Wheat phytase treatment was found more effective for PA, whereas, the Aspergillus phytase was found more effective in increasing the inorganic P content in treated bran samples. The inorganic P content in phytase treated bran sample was increased in the range of 16.30-29.62 % over untreated with maximum content was obtained in bran of WH 711 &WH 1021. Protein content after phytase treatment was increased for crude protein, whereas, soluble protein content was reduced to different extent. It was revealed that under given experimental conditions, phytase treatment resulted in decreased micronutrient dialyzability to different extent, with 7.11-45% reduction with wheat phytase treatment. Overall, wheat phytase treatment was found as comparatively better over Aspergillus phytase under given experimental conditions and bran of WH 1021 was found superior in studied quality attributes under given treatments. It is proposed to conduct the similar study with higher dose of phytase enzyme and in vitro digestion similar to gastric conditions before dialysis could give a greater and better insight into this area. The decreased dialyzability might be associated to experimental conditions and lower phytase dose. However, the results of the study showed that adding phytase to wheat bran improved its nutritional value for food applications.ThesisItem Open Access Effect of exogenous melatonin on cotton seed germination and seedling under saline condition(CCSHAU, Hisar, 2021-09) Jakhar, Sonika; Mandhania, ShiwaniCotton is an important fiber crop grown throughout the world and contributes significantly to global economy. Salinity is most destructive abiotic stress particularly at seed germination and seedling stage which considerably affects cotton productivity. Melatonin is regarded as a plant growth-promoting molecule and helps in mitigating several different abiotic stresses including salinity. Therefore, the present investigation was carried to study the effect of melatonin on seed germination and biochemical constituents in cotton under salinity stress. Results showed that salinity causes negative effects on cotton seed germination and decreases seed vigor. It also enhanced relative stress injury (in terms of electrolyte leakage), H2O2 and MDA content of seedlings by increasing membrane permeability to ions and solutes. Exogenous application of melatonin at the concentration of 20 μM has profound effect on seed germination, seedling vigor traits and antioxidant defense system in cotton under salinity soil. This indicates that seed priming with 20 μM melatonin mitigates the salinity effect and significantly improves cotton seed germination and vigor index traits. Melatonin might improve cotton germination under saline stress by mitigating the negative effect of oxidative stresses through elevating antioxidant defense system. This study provides an important information to improve cotton productivity in salt affected soil.ThesisItem Open Access Biochemical studies on antioxidative defense system in advance lines of wheat (Triticum aestivum L. em Thell.) grown under salt stress(CCSHAU, Hisar, 2021-08) Bhadu, Surina; Tokas, JayantiWheat (Triticum aestivum) is an important cereal crop for the world population. Based on productivity, wheat is the third most commonly grown cereal after maize and rice. It is essential for providing the mandatory amount of calories and protein content in the affluent human diet. Among abiotic stresses, salt stress is the chief reason which affects approximately 50% of crop productivity along with soil fertility. Under saline conditions, there is an imbalance in the uptake of the ions which disturbs the homeostasis between Na+/ K+ ratio. Due to the ions imbalance in plants, there is a generation of ionic and osmotic stress at the cellular level. The excess amount of ROS produced is the consequence of this stress which damages the membrane integrity that causes several changes at physiological, morphological, biochemical, and molecular levels. The present study was carried out on F5 and F6 generation of a conventional wheat cross between Kharchia 65 (salt-tolerant) × WH1105 (saltsensitive). The F5 generation was sown at 8 ECe while F6 was sown at 10 ECe and analyses were carried out at vegetative and reproductive stages. Under salt stress, WH1105 had higher oxidative stress components viz. H2O2, MDA which increased in response to the ROS and antioxidative defense system got activated to scavenges the excess of ROS. Kharchia 65 (salt-tolerant) showed higher antioxidative enzyme activities than WH1105 (salt-sensitive) under salt stress at both vegetative and reproductive stages. Introgress of Nax1 and Nax2 genes was much affective in increasing the antioxidative enzyme activities viz. at vegetative stage, SOD (1.33%, 4.50%), CAT(14.6%, 4.13%), APX(12.57%, 20.64%), GR (14.51%, 20.34%), GPX(13.91%, 4.34%), MDHAR (55.8%, 44.4%), DHAR (42.9%, 38.3%) and POX (10.44%, 4.29%) and at reproductive stage, SOD (3.26%, 3.25%), CAT(4.67%, 7.18%), APX(11.87%, 9.13%), GR (2.65%, 19.8%), GPX(19.16%, 8.48%), MDHAR (45.7%, 48.6%), DHAR (35.7%, 34.2%) and POX (7.58%, 7.12%) in F5 and F6 generations respectively in plants expressing Nax1 and Nax2 genes as compared to Kharchia 65 under salt stressed condition. The physiological parameter like RWC and osmotic potential declined with an increase in salinity and the Na+/K+ ratio was higher in F6 as compared to F5. The LOX activity was also higher in WH1105 indicating higher lipid peroxidation as compared to Kharchia 65. The proteomics data indicate that there is up-regulation of many proteins in the plants that expressing either Nax1 or Nax2 gene as compared to salt stressed Kharchia 65. From the present study, it is concluded that introgress of Nax1 and Nax2 genes had activated the antioxidative defense mechanism with an increase in the crop yield under salt stress.ThesisItem Open Access Biochemical and expression profiling of sorghum [Sorghum bicolor (L.) Moench] under salt stress(CCSHAU, Hisar, 2021-04) Himani; Tokas, JayantiSalinity induces complex metabolic processes that involve ion toxicity, osmotic stress, biochemical and physiological perturbations. Sorghum is a gluten-free cereal crop, well adapted to semi-arid tropics, highly biomass productive, and water-efficient. Therefore, the present study aimed to assess sorghum genotypes' tolerance behavior under different salinity levels (6, 8, 10, and 12 dSm-1) concerning the performance of physiological, biochemical, and molecular responses. Amongst 23 screened sorghum genotypes, SSG59-3 was identified as salt-tolerant and PC-5 as salt susceptible genotype based on germination studies. The results revealed that at 35 and 95 DAS, morpho-physiological parameters (RWC, OP, CF, CSI, total chlorophyll content) and growth traits (plant height, fresh and dry weight, root and shoot length) decreased with increasing salt concentration. The maximum decrease was observed in PC-5 as compared to SSG 59-3 at 10 dSm-1. PC-5 accumulated higher Na+and K+ content in roots while the exclusion potential of SSG 59-3 was more in roots, so it had minimum Na+ and K+ content. The quality traits viz. HCN, CP, IVDMD, CPY, DDM, and cell wall components (except HC, which decreased with plant maturity) also followed a similar pattern. SSG 59-3 maintained the higher GFY, DMY, and yield components at 10 dSm-1 (at physiological maturity). The imposition of salinity induces the activities of ROS scavenging enzymes viz. SOD, CAT, POX, APX, GPX, GR, MDHAR, and DHAR and the content of ROS scavenging metabolites viz. ascorbate, glutathione, and carotenoids which were maximum in SSG 59-3 while lower in PC-5 at 10 dSm-1. The oxidative stress as judged by accumulation of MDA and H2O2 content was maximum in PC-5 and minimum in SSG 59-3. A significant increase was observed in compatible osmolytes viz. proline, glycine betaine, and total carbohydrates, which was more pronounced in SSG 59-3 than PC-5 at 10 dSm-1. Analysis of the LFQ results using MASCOT software revealed more than 100 differentially expressed proteins, out of which 40 proteins were upregulated, 29 proteins were down regulated and the majority was involved in catalytic activity, binding proteins, metabolic inter-conversion enzymes and ion transporters. The relative expression of CIPK24, LEA3, BADH1 was highly upregulated at 10 dS m-1while NCED3, SNAC1, GPX, H+-PPase, and P5CS1 were up-regulated at 12 dS m-1. The fold change was maximum in SSG 59-3 while the expression level of transcripts was less in PC-5. Hence, SSG 59-3 seems physiologically and biochemically promising and may be exploited in plant breeding programs aimed at developing salt-tolerant sorghum genotypes.
